STRUCTURE DETERMINATION OF THE SAR/SEC23/PEPTIDE

SAR/SEC23/肽的结构测定

• 批准号: 7726223
• 负责人: JONATHAN D GOLDBERG
• 金额: $ 0.52万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-18 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7726223
• 关键词: Binding; Clathrin; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; Endoplasmic Reticulum; Funding; Golgi Apparatus; Grant; Image; Institution; Manuscripts; Membrane; Molecular; Peptides; Proteins; Publishing; Research; Research Personnel; Resources; SNAP receptor; Series; Source; Structure; Transport Vesicles; United States National Institutes of Health; Yeasts; beamline; interest

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This is the third in a series of proposals that aims at obtaining structural information on the yeast COPII vesicular coat. COPII is a 420 kDa pentameric complex responsible for budding transport vesicles from the endoplasmic reticulum (ER) and delivering the specific cargo contents to the Golgi complex. In the first proposal we focused on the inner shell of the COPII coat, the Sec23/24-Sar1 complex. We were able to solve this 210 kDa structure by MAD and molecular replacement from data collected at X-25 and at CHESS (manuscript published). The second proposal focused on the selectivity of the COPII coat for cargo and for ER-to-Golgi SNARE fusogenic proteins. We were able to determine the structures of three complexes of Sec24 bound to cargo and SNARE sequences (manuscript submitted). In the current proposal we focus on the outer shell of COPII, comprising the Sec13/31 complex. Sec13/31 is functionally (but not structurally) homologous to clathrin-it polymerizes the COPII inner shell on the ER membrane and causes membrane deformation. Since, like clathrin, Sec13/31 is a long, flexible structure capable of oligomerizing (as seen in EM images) it is unlikely that the intact complex can be crystallized. Therefore, we have dissected the protein into what we believe are the two functionally-interesting fragments, and these constitute the two projects that we propose in this beamline application.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 这是旨在获取酵母 COPII 囊泡外壳结构信息的一系列提案中的第三个提案。 COPII 是一种 420 kDa 五聚体复合物，负责从内质网 (ER) 出芽运输囊泡，并将特定的货物内容物输送到高尔基复合体。在第一个提案中，我们重点关注 COPII 涂层的内壳，即 Sec23/24-Sar1 复合体。我们能够通过 MAD 和分子替换从 X-25 和 CHESS 收集的数据中解析出这个 210 kDa 的结构（手稿已发表）。第二个提案侧重于 COPII 涂层对货物和 ER-to-Golgi SNARE 融合蛋白的选择性。我们能够确定与货物和 SNARE 序列结合的 Sec24 的三种复合物的结构（已提交手稿）。 在当前的提案中，我们重点关注 COPII 的外壳，包括 Sec13/31 复合体。 Sec13/31在功能上（但在结构上）与网格蛋白同源——它聚合ER膜上的COPII内壳并导致膜变形。由于与网格蛋白一样，Sec13/31 是一种长而灵活的结构，能够寡聚（如 EM 图像中所示），因此完整的复合物不太可能结晶。因此，我们将蛋白质解剖成我们认为的两个功能上有趣的片段，这些片段构成了我们在此光束线应用中提出的两个项目。